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Cambridge International Examinations

Cambridge International Advanced Subsidiary and Advanced Level


* 7 3 5 1 7 5 0 3 6 6 *

BIOLOGY 9700/33
Paper 3 Advanced Practical Skills 1 February/March 2018
2 hours
Candidates answer on the Question Paper.
Additional Materials: As listed in the Confidential Instructions.

READ THESE INSTRUCTIONS FIRST

Write your Centre number, candidate number and name on all the work you hand in.
Write in dark blue or black pen.
You may use an HB pencil for any diagrams or graphs.
Do not use staples, paper clips, glue or correction fluid.
DO NOT WRITE IN ANY BARCODES.

Answer all questions.

Electronic calculators may be used.


You may lose marks if you do not show your working or if you do not use appropriate units.

At the end of the examination, fasten all your work securely together.
The number of marks is given in brackets [ ] at the end of each question or part question.

For Examiner’s Use

Total

This document consists of 14 printed pages and 2 blank pages.

DC (ST/SW) 145387/4
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Before you proceed, read carefully through the whole of Question 1 and Question 2.

Plan the use of the two hours to make sure that you finish all the work that you would like to do.

If you have enough time, think about how you can improve the confidence in your results, for
example by obtaining and recording one or more additional measurements.

You will gain marks for recording your results according to the instructions.

1 Mung bean seeds contain an enzyme that is used to hydrolyse (break down) sucrose into reducing
sugars. This enzyme is essential to provide the reducing sugars needed for the seeds to grow.

When mung bean seeds are soaked in sucrose solution, some of this enzyme diffuses into the
surrounding solution and hydrolyses the sucrose.

enzyme from seeds


sucrose glucose and fructose

The apparatus was set up as shown in Fig. 1.1.

beaker

sucrose solution

mung bean seeds

Fig. 1.1

Samples of the sucrose solution were removed at 10 minutes (sample S1), at 15 minutes
(sample S2) and at 30 minutes (sample S3) after adding the sucrose solution.

You are required to:

• make a serial dilution of 1.0% reducing sugar solution, R


• carry out the Benedict’s test on each concentration of reducing sugar
• carry out the Benedict’s test on S1, S2 and S3
• estimate the concentration of reducing sugar in S1, S2 and S3.

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You are provided with the materials shown in Table 1.1.

Table 1.1

labelled contents hazard volume / cm3


R 1.0% reducing sugar solution none 50
S1 sample removed after 10 minutes none 20
S2 sample removed after 15 minutes none 20
S3 sample removed after 30 minutes none 20
W distilled water none 100
Benedict’s Benedict’s solution harmful 40

It is recommended that you wear suitable eye protection. If Benedict’s comes into contact with
your skin, wash it off immediately under cold water.

1. Set up a water-bath and heat the water to a suitable temperature to test for reducing sugars
using the Benedict’s test.

(a) (i) State the temperature you will need to maintain in the water-bath to carry out the
Benedict’s test.

temperature ...........................................................[1]

You are required to make a serial dilution of the 1.0% reducing sugar solution, R, which
reduces the concentration by half between each successive dilution. This will provide you
with a set of reducing sugar solutions of known concentrations.

After the serial dilution is completed, you will need to have 10 cm3 of each concentration
available for use.

Fig. 1.2 shows the first two beakers that you will use to make your serial dilution.

(ii) Complete Fig. 1.2 by drawing as many extra beakers and arrows as you need to show
how you will carry out your serial dilution.

For each beaker:

• state, under the beaker, the volume and concentration of the reducing sugar
solution in the beaker that will be available for use in the investigation, after the serial
dilution has been completed

• use one arrow, with a label above the beaker, to show the volume and concentration
of reducing sugar solution added to prepare the concentration of the reducing sugar
solution in the beaker

• use another arrow, with a label above the beaker, to show the volume of distilled
water, W, added to prepare the concentration of reducing sugar solution in the beaker.

The first part of Fig. 1.2 has been labelled for you.

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20 cm3 of 1.0%
.......................
reducing sugar
.......................
.......................
solution, R
.......................
.......................
0 cm3
of W .......................

10 cm3 of 1.0%
.......................
reducing sugar
.......................
solution to use
.......................
.......................

.......................

.......................

Fig. 1.2

[3]

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Read step 2 to step 4 before proceeding.

2. Prepare all the concentrations of reducing sugar solution shown in Fig. 1.2, in the beakers
provided.

(iii) You will need to carry out a Benedict’s test on each of the different concentrations of
reducing sugar solution and on 2 cm3 of each of S1, S2 and S3.

You will be recording the time taken for the first appearance of a colour change.

State the volume of Benedict’s solution and the volume of each of the concentrations of
reducing sugar solution you will use for each test.

volume of Benedict’s solution ...............................................................

volume of each concentration of reducing sugar solution ...............................................................


[1]

3. Using the volumes you decided in (a)(iii), carry out the Benedict’s test on the reducing sugar
solutions of different concentrations shown in Fig. 1.2.

Test one solution at a time, using the syringe labelled B for the Benedict’s solution.

Record, in (a)(iv), the time taken for the first appearance of a colour change.

If there is no colour change after 120 seconds, record as ‘more than 120’.

(iv) Record your results in an appropriate table.

[4]
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You are required to estimate the concentration of reducing sugars in S1, S2 and S3.

4. Carry out the Benedict’s test on 2 cm3 of each of S1, S2 and S3 and record the time taken for
the appearance of the first colour change.

(v) State the time taken for the appearance of the first colour change for S1, S2 and S3.

S1 .......................... S2 .......................... S3 .......................... [1]

(vi) Complete Fig. 1.3 by:

• labelling the position on the line of each of the percentage concentrations of reducing
sugar solution shown in Fig. 1.2

• putting the labels S1, S2 and S3 on Fig. 1.3 to show an estimate of the concentrations
of reducing sugar in S1, S2 and S3.

0.0% 1.0%

percentage concentration of reducing sugar

Fig. 1.3
[2]

(vii) Sample S1 was removed from the sucrose solution 10 minutes after seeds had been
added. Sample S2 was removed 15 minutes after the seeds had been added.

Suggest an explanation for the difference in results for S1 and S2.

...........................................................................................................................................

.......................................................................................................................................[1]

(viii) If the mung bean seeds are soaked in the sucrose solution for more than 30 minutes, the
concentration of reducing sugar remains the same as S3.

Use your knowledge of enzymes to explain this observation.

...........................................................................................................................................

.......................................................................................................................................[1]

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(b) A student set up the apparatus shown in Fig. 1.4 to investigate whether a different type of
seed also releases an enzyme that hydrolyses sucrose.

beaker

distilled water

seeds

Fig. 1.4

After 30 minutes, the student tested the water for protein. This test showed that protein was
present in the water.

(i) State the name of the test for protein.

.......................................................................................................................................[1]

The student suggested the hypothesis:

the protein in the water is an enzyme that hydrolyses sucrose.

(ii) The student mixed 2 cm3 of the water containing the protein with 5 cm3 of 1% sucrose
solution. After 30 minutes, reducing sugars were present.

Describe how the student could have set up a suitable control for this experiment to
provide evidence that hydrolysis of sucrose was due to an enzyme.

...........................................................................................................................................

...........................................................................................................................................

...........................................................................................................................................

...........................................................................................................................................

.......................................................................................................................................[2]

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Another student tested the concentration of this enzyme that had been released from the seeds of
several different species using a standard method.

The results are shown in Table 1.2.

Table 1.2

concentration of
species of
enzyme
plant
/ arbitrary units
F 9.5
G 15.0
H 17.0
J 20.5
K 39.5

(c) Draw a bar chart of the data in Table 1.2 on the grid in Fig. 1.5.

Each bar should be separated for each species of plant.

Use a sharp pencil for drawing bar charts.

Fig. 1.5

[4]

[Total: 21]

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2 (a) P1 is a slide of a stained transverse section through a plant leaf.

You are not expected to be familiar with this specimen.

You are required to:

• use the eyepiece graticule to measure the depth of the leaf and the depth of a vascular
bundle at the mid-rib
• use these measurements to draw a plan diagram of part of the leaf.

The eyepiece graticule in the microscope can be used to measure different tissues.

Select the widest part of the leaf (mid-rib) on P1, shown by Y in Fig. 2.1.

Fig. 2.1

(i) Use the eyepiece graticule in the microscope to measure:

• the depth of the leaf at the mid-rib


• the depth of the vascular bundle at the mid-rib.

depth of leaf ........................ eyepiece graticule units

depth of vascular bundle ........................ eyepiece graticule units


[1]

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Use a sharp pencil for drawings.

(ii) Use the measurements from (a)(i) to help you to draw a large plan diagram of the section
of the leaf shown by the shaded area in Fig. 2.2.

Use one ruled label line and label to identify the vascular bundle.

draw this
section
upper surface

lower surface

Fig. 2.2

You are expected to draw the correct shape and proportions of the different tissues.

[5]

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Question 2 continues on page 13

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(iii) Observe the upper epidermis of the leaf and the layer of cells below the upper epidermis.
Fig. 2.2 identifies the upper and lower surfaces of the leaf.

Select three epidermal cells and one cell in the layer below that touches these cells.

Make a large drawing of this group of four cells. Each cell must touch at least one of the
other cells. Do not draw the cuticle.

Use one ruled label line and label to identify the cell wall of one cell.

[5]

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(b) Fig. 2.3 is a photomicrograph of a stained transverse section through a leaf of a different type
of plant.

You are not expected to be familiar with this specimen.

L2

L3

L1

magnification ×40

Fig. 2.3

(i) Determine the simplest whole number ratio of the total length of L1 and L2 (the leaf
tissue) to the length of L3 (the air space).

Show all the steps in your working.

simplest whole number ratio ...............................................................


[4]

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(ii) Observe the leaf on slide P1 and the leaf shown in Fig. 2.3.

Identify one feature to reduce water loss that can be observed for both leaves.

Describe how this feature reduces water loss.

feature ...............................................................................................................................

description .........................................................................................................................

...........................................................................................................................................
[1]

(iii) Observe the leaf on P1 and the leaf in Fig. 2.3 and identify differences between them.

Record the observable differences in Table 2.1.

Table 2.1

feature P1 Fig. 2.3

[3]

[Total: 19]

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